摘要
目的 构建人疱疹病毒7型(HHV-7)开放读码框(ORF)U14的原核表达载体,并且进行原核表达。方法 PCR技术扩增HHV-7 ORF U14基因的主要抗原决定簇编码区(328~533氨基酸),目的基因与原核表达载体pThioHis A分别双酶切后连接,1×TSS法转化宿主菌E.coli BL21,异丙基-β-D-硫代半乳糖苷诱导融合蛋白表达。结果 基因序列分析表明目的基因序列与HHV-7标准毒株相应序列基本-致,SDS-PAGE电泳可观察到相对分子质量为3.57万融合蛋白的表达,Western印迹证实pp85蛋白成功表达。结论 HHV-7被膜蛋白pp85原核表达载体构建和表达成功,为进一步探讨该重组蛋白在HHV-7特异性抗体检测中的应用提供实验基础。
Objective To construct the prokaryotic expression vector of tegument protein pp85 of human herpesvirus 7 and to express it. Methods The major antigen epitopes coding sequence (328-533 aa) of HHV70RF U14 gene was amplified by PCR and inserted into the prokaryotic expression vector pThioHis A by gene engineering technique; recombinant plasmid was transformed into E. coil BL21 and expressed by IPTG inducing. Results The target gene was identical with that of HHV-7 standard species,and the 35 700 fusion protein was found in SDS-PAGE and proved by western blot. Conclusion Recombinant prokaryotic expression vector was gained, which is valuable for the further study on HHV-7 specific antibody detection.
出处
《青岛大学医学院学报》
CAS
2006年第1期47-49,52,共4页
Acta Academiae Medicinae Qingdao Universitatis
基金
青岛市科技局资助项目(2001KNS-2E-27-1)